|
| Status |
Public on Aug 09, 2023 |
| Title |
Renal tissue of the onilateral ureteral ligation (UUO) mice (model) 1 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Renal cortex of UUO mice
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
age: 8 week
gender: male
type: IP enriched m6A methylated RNAs
|
| Treatment protocol |
For the unilateral ureteral ligation-induced UUO kidney fibrosis model, mice received permanent ligation of the free ureter in the left kidney under aseptic and anesthetic conditions, while sham-operated mice received the free ureter but not ligation under equivalent conditions. Seven days after surgery, mice were sacrificed under anesthesia to obtain renal cortical tissue.
|
| Growth protocol |
Mice were routinely reared to 22-24 g after they reached 8 weeks of age.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNAs were immunoprecipitated with anti-N6-methyladenosine(m6A) antibody. The immunoprecipitated "IP" fraction contained enriched m6A methylated RNAs, and the supernatant " Sup" fraction contained unmodified RNAs.
|
| Label |
cy5
|
| Label protocol |
"IP" and "Sup" RNAs were amplified as cRNAs and labeled with Cy5 and Cy3 respectively using Arraystar Super RNA Labeling Kit.
|
| |
|
| Channel 2 |
| Source name |
Renal cortex of UUO mice
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
age: 8 week
gender: male
type: supernatant unmodified RNAs
|
| Treatment protocol |
For the unilateral ureteral ligation-induced UUO kidney fibrosis model, mice received permanent ligation of the free ureter in the left kidney under aseptic and anesthetic conditions, while sham-operated mice received the free ureter but not ligation under equivalent conditions. Seven days after surgery, mice were sacrificed under anesthesia to obtain renal cortical tissue.
|
| Growth protocol |
Mice were routinely reared to 22-24 g after they reached 8 weeks of age.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNAs were immunoprecipitated with anti-N6-methyladenosine(m6A) antibody. The immunoprecipitated "IP" fraction contained enriched m6A methylated RNAs, and the supernatant " Sup" fraction contained unmodified RNAs.
|
| Label |
cy3
|
| Label protocol |
"IP" and "Sup" RNAs were amplified as cRNAs and labeled with Cy5 and Cy3 respectively using Arraystar Super RNA Labeling Kit.
|
| |
|
| |
| Hybridization protocol |
Cy3 and Cy5 labeled cRNAs were combined together and hybridized to Arraystar Mouse mRNA&lncRNA Epitranscriptomic Arrays (8x60K, Arraystar) at 65°C for 17 hours in an Agilent Hybridization Oven.
|
| Scan protocol |
After washing, slides were scanned with Agilent Scanner G2505C.
|
| Data processing |
Agilent Feature Extraction software (version 11.0.1.1) was used to analyze acquired array images. Raw intensities of IP (immunoprecipitated, Cy5-labelled) and Sup (supernatant, Cy3-labelled) were normalized with average of log2-scaled Spike-in RNA intensities. After Spike-in normalization, the probe signals having Present (P) or Marginal (M) QC flags in at least 3 out of 6 samples were retained for further “m6A quantity” analyses. “m6A quantity” was calculated for the m6A methylation amount based on the IP (Cy5-labelled) normalized intensities.
|
| |
|
| Submission date |
Mar 02, 2023 |
| Last update date |
Aug 09, 2023 |
| Contact name |
Wei-Jian Ni |
| Organization name |
University of Science and Technology of China
|
| Department |
Anhui Provincial Hospital, The First Affiliated Hospital of USTC
|
| Lab |
Department of Pharmacy
|
| Street address |
No. 17 Lujiang Road
|
| City |
Hefei |
| State/province |
Anhui |
| ZIP/Postal code |
230001 |
| Country |
China |
| |
|
| Platform ID |
GPL25915 |
| Series (1) |
| GSE226505 |
Mouse kidney tissue: Control vs. UUO |
|
